Determination of 3′,4′,5′,5,7-pentamethoxyflavone in the plasma and intestinal mucosa of mice by HPLC with UV detection

Abstract

This paper was published as Biomedical Chromatography, 2009, 23 (4), pp. 335-339. It is available from http://onlinelibrary.wiley.com/doi/10.1002/bmc.1120/abstract. DOI: 10.1002/bmc.1120Metadata only entryIn a preliminary experiment 3′,4′,5′,5,7-pentamethoxyflavone (PMF) inhibited adenoma development in ApcMin mice, a model of the human heritable condition familial adenomatous polyposis. An HPLC method for tricin was modified and validated to permit measurement of PMF in mouse plasma and intestinal mucosa. HPLC analysis was carried out on a Hypersil-BDS C18 column with detection at 324 nm and tricin as internal standard. The assay was linear in the range of 100–2000 ng/mL plasma and 1.0–40 µg/mL mucosa. PMF in plasma was efficiently extracted using solid-phase columns. In the case of mucosa organic solvent protein precipitation displayed satisfactory accuracy and precision. The assay recovery at low, medium and high concentrations was between 85 and 103% for both biomatrices, with a relative standard deviation of <15%. The lower limits of quantitation for plasma and mucosa were 100 ng/mL and 1.0 µg/mL, respectively. This method allowed measurement of PMF steady-state median concentrations in plasma (1.08 nmol/mL, n = 11; 10th and 90th percentiles: 0.633 and 2.385 nmol/mL) and mucosa (108.5 nmol/g, n = 9; 10th and 90th percentiles: 38.9 and 164.4 nmol/g) in mice which had received PMF (0.2%, w/w) with their diet